Is the big cell a principal open set?

This is true if $G$ is (semi-simple) simply-connected, because then $\mathrm{Pic}(G)=(0)$, which means that $\mathbb{C}[G]$ is factorial; however, it is false for the simplest non simply-connected example, namely $G=\mathrm{PGL}(2)$. Indeed $G$ is the complement of the quadric $ad-bc=0$ in $\mathbb{P}^3$; this implies that $\mathrm{Pic}(G)=\mathbb{Z}/2$, generated by the line bundle $\mathcal{O}_{\mathbb{P}^3}(1)$ restricted to $G$. The complement of the big cell is the divisor $a=0$, whose class in $\mathrm{Pic}(G)$ is the nonzero element; hence it is not principal.